• Title, Summary, Keyword: Respiration inhibition assay

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Efficient assay for respiration inhibitor using Saccharomyces cerevisiae (Saccharomyces cerevisiae를 이용한 효율적인 호흡저해제 검정법)

  • Choi, Gyung-Ja;Kim, Jin-Cheol;Kim, Heung-Tae;Cho, Kwang-Yun
    • The Korean Journal of Pesticide Science
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    • v.4 no.3
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    • pp.52-59
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    • 2000
  • A rapid assay to determine respiration inhibition of Saccharomyces cerevisiae by chemicals was developed. S. cerevisiae was harvested with two different liquid media, yeast extract-peptone-dextrose (YPD) medium capable of occurring both glucose fermentation and mitochondrial respiration, and non-fermentable carbon-yeast extract (NFY) medium capable of occurring respiration only Wells in 96-well plate were loaded with each cell suspension and various concentrations of 46 fungicides with various modes of action. n NFY medium, the non-fermentable carbon source, ethanol (NFY-E medium), glycerol (NFY-G medium) or lactate (NFY-L medium), was used. After incubation for $1{\sim}3$ days, minimum inhibitory concentrations (MICs) of the chemicals were recorded in the media. Of the 46 inhibitors employed in this study, four inhibitors of fungal respiration by blockage of electron flux in the mitochondrial respiratory chain, azoxystrobin, kresoxim-methyl, metominostrobin, and trifloxystrobin, exhibited strong antifungal activity in all of NFY media, but no activity in YPD medium. In contrast to this, five N-trihalomethylthio fungicides showed much stronger antifungal activities in YPD medium than three NFY media. Eleven fungicides inhibited growth of S. cerevisiae in all media and the other 26 fungicides showed no antifungal activity in all media. Thus, our rapid and efficient in vitro method can be considered as an alternative assay system for respiration inhibitor.

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Blue Light Photosensitization in Mitochondrial Membrane of Plant Cells (식물세포 미토콘드리아막에서 일어나는 청색광 Photosensitization)

  • Kim, Kyung-Hyun;Kim, Jong-Pyung;Jung, Jin
    • Korean Journal of Environmental Agriculture
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    • v.6 no.2
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    • pp.94-100
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    • 1987
  • Plant mitochondria, irradiated with blue-colored $sunlight(350{\sim}500nm)$ under aerobic and anaerobic conditions, were assayed as to the electron transfer activity of respiratory enzyme system, and compared with those irradiated with orange-colored light(white sunlight minus blue-colored light). The respiratory activity of mitochondria was most seriousely inhibited by illumination with blue-colored light under aerobic condition. Deaeration of mitochondrial suspension resulted in substantial decrease of the photoinhibition by blue-colored light. Meanwhile, orange-colored light demonstrated much less effectiveness-almost ineffectiveness-in causing the inhibition of mitochondrial respiration system. The results of enzymatic assay revealed a strong possibility that FMN in NDH and heme group at least in cytochrome c oxidase, but not FAD in SDH, are the photodynamic sensitizers in mitochondrial inner membrane. Also worthwhile to note is the significant difference from the others of SDH in its photoinhibitory response to the light quality of visible light; that the inhibition of SDH by irradiation was not affected by atmospheric condition and that orange-colored light gave rise to considerable extents of inhibition to the enzyme. This observation was tentatively interpreted in terms of photosensitized reaction not involving molecular oxygen possibly catalyzed by Fe-S centers in the enzyme. The superoxide production and the membrane peroxidation of mitochondria under various treatments also indicated that there was blue-light photodynamic reaction in mitochondria involving active oxygens.

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